The Relationship Between CLSPN Expression And Prognosis In Lung Adenocarcinoma

Objective:Lung cancer is the leading cause of cancer-associated mortality in the world.Lung cancer is mainly divided into non-small cell lung cancer(NSCLC)and small cell lung cancer(SCLC).Non-small cell lung cancer accounts for about 85%.lung adenocarcinoma(LUAD)is a major type of non-small cell lung cancer,accounting for about 40%.The treatment of NSCLC has made encouraging progress over the past few decades and primarily consists of surgical resection,chemotherapy,immunotherapy and molecular targeting treatment.However patients often have distant metastasis when diagnosed or experience recurrence after surgery,so only 17.4% patients survive longer than 5 years.Therefore,it is important to explore possible targets to improve the prognosis of patients with lung adenocarcinoma.The purpose of this study was to evaluate the expression of CLSPN in lung adenocarcinoma and its effect on the proliferation,migration and invasion of lung cancer cells.The gene might be a potential prognostic and diagnostic biomarker for patients with LAUD.In addition,CLSPN inhibitors may serve as effective therapies for LAUD,especially the patients with distant metastases.Methods:1.We downloaded clinical data which contained the prognosis and basic clinical information and the sequencing data of mRNA of 513 LAUD samples and 59 normal samples from the Cancer Genome Atlas(TCGA,https://portal.gdc.cancer.gov/).2.Lung adenocarcinoma cells A549 and PC-9 cells were cultured at 37? in 5% CO2 fetal bovine serum medium to maintain good cell growth.Then we add the optimal amount of virus for infection,and it can continue to be used for in vivo and in vitro research.We generally choose two cell lines with higher infection rates for downstream experiments.3.Based on the first two steps,CCK8 method,flow cytometry,scratch test,and transwell invasion experiment were used to investigate the effect of claspin expression level on the biological behavior of lung adenocarcinoma cells in vitro after lung adenocarcinoma was infected with CLSPN shRNA.Results:1.By analyzing the clinical data of patients in the database,the expression of CLSPN in lung adenocarcinoma cells is significantly higher than that of cells in adjacent lung tissues in humans,and the expression of CLSPN in patients with advanced TNM stage is higher than that in early patients More significantly,especially in patients with extrapulmonary metastases,CLSPN increased significantly in cancer cells.We found that the patients with high CLSPN expression were younger,but the mortality was higher.COX regression found that the expression of CLSPN was indeed related to the survival time of the patients,and it is obvious from the survival curve that the patients with high CLSPN expression survive The time is shorter,so the level of CLSPN expression is an independent risk factor for the prognosis of patients,which has initially suggested the cancer-promoting effect of this gene.2.We selected human adenocarcinoma cells A549 and PC-9 cells for real-time quantitative PCR.The results showed that the gene CLSPN was abundantly expressed in both types of cell beads.CLSPN expression was significantly suppressed in A549 and PC-9 cell lines treated with CLSPN shRNA(P <0.05),thus confirming the knockdown phenomenon.We selected PC-9 cell lines with higher knockdown rates for further research.3.The proliferation of lung adenocarcinoma cell lines treated with CLSPN shRNA was significantly slowed,but apoptosis was significantly increased.In addition,the cell migration and invasion ability was significantly inhibited after virus transfection.Our in vitro experiments proved that the expression of CLSPN can be effectively suppressed at the cellular level,and further confirmed that the gene promotes the development of lung adenocarcinoma.Conclusions:Compared with adjacent tissues,the expression of CLSPN in lung cancer tissues was significantly increased(p <0.05).The high expression of CLSPN in lung cancer tissues is significantly related to the late stage of pathological TNM staging and distant metastasis,but not related to gender,race,tumorigenesis site,and leaf segments.CLSPN overexpression is associated with poor prognosis in patients with lung adenocarcinoma and is an independent prognostic factor for patients with lung adenocarcinoma.Therefore,CLSPN can promote the proliferation,migration and invasion of lung cancer cells.Therefore,further studying the mechanism of this gene and applying it to clinical practice can bring new directions and hope for the treatment of patients with advanced lung adenocarcinoma.